Development in Wastewater Treatment Research and Processes: Microbial Degradation of Xenobiotics Through Bacterial and Fungal Approach Maulin P. Shah

Development in Wastewater Treatment
Research and Processes: Microbial

Degradation of Xenobiotics through
Bacterial and Fungal Approach Maulin
P. Shah
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Development in
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Research and
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Development in
Wastewater Treatment
Research and
Processes
Microbial Degradation of
Xenobiotics Through Bacterial
and Fungal Approach
Edited by
Susana Rodriguez-Couto
Department of Separation Science, LUT School of
Engineering Science, LUT University, Mikkeli, Finland
Maulin P. Shah
Environmental Microbiology Lab, Bharuch, Gujarat, India
Elsevier
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Contents
Contributors ....................................................................................................................................... xvii
CHAPTER 1 Microbial degradation of xenobiotics in bioelectrochemical

systems.................................................................................................. 1
Somdipta Bagchi and Manaswini Behera
1.1 Introduction to xenobiotics .........................................................................................1
1.2 Conventional processes of xenobiotics degradation and their drawbacks.................3
1.2.1 Adsorption ........................................................................................................ 3
1.2.2 Membrane processes ........................................................................................ 4
1.2.3 Advanced oxidation processes ......................................................................... 4
1.2.4 Biological treatment processes......................................................................... 5
1.3 Xenobiotics degradation in bioelectrochemical systems ...........................................6
1.3.1 Microbial fuel cell ............................................................................................ 6
1.3.2 Microbial electrolysis cell .............................................................................. 11
1.3.3 Microbial desalination cell............................................................................. 14
1.4 Challenges faced .......................................................................................................15
1.5 Summary ...................................................................................................................15
1.6 Conclusion ................................................................................................................16
References.................................................................................................................16
Further reading .........................................................................................................22
CHAPTER 2 Bacterial and fungal degradation of dyes: a remedial source ......... 23
Bhumika Rajoria and Arpita Roy
2.1 Introduction.............................................................................................................23
2.2 Classification of dyes..............................................................................................24
2.3 Environmental and health problems associated with dyes ....................................25
2.4 Conventional methods of dye removal...................................................................26
2.5 Bacterial degradation of dye ..................................................................................28
2.5.1 Pure bacterial cultures for dye degradation .................................................28
2.5.2 Mixed cultures and cocultures for degradation ...........................................29
2.5.3 Immobilized cells for degradation ...............................................................29
2.5.4 Microbial fuel cells for degradation ............................................................30
2.6 Mechanism of dye degradation via bacteria ..........................................................30
2.6.1 Degradation via enzymes .............................................................................30
2.6.2 Degradation via mediation ...........................................................................32
2.6.3 Decolorization via organic and inorganic compounds ................................32
2.7 Factors affecting bacterial degradation of dyes .....................................................32
2.7.1 Aeration and agitation ..................................................................................32
v
vi Contents
2.7.2 Temperature ..................................................................................................33

2.7.3 pH .................................................................................................................33
2.7.4 Dye concentration.........................................................................................33
2.7.5 Dye structure ................................................................................................33
2.8 Bacterial bioreactors for decolorization and degradation of dyes.........................34
2.9 Fungal degradation of dye......................................................................................34
2.10 Mechanism of dye degradation via fungi...............................................................35
2.10.1 Bioaccumulation of dye .............................................................................35
2.10.2 Biosorption of dye......................................................................................36
2.10.3 Biodegradation of dye ................................................................................36
2.11 Factors affecting fungal degradation......................................................................37
2.11.1 Aeration and agitation ................................................................................37
2.11.2 Temperature ................................................................................................37
2.11.3 pH ...............................................................................................................38
2.11.4 Dye concentration.......................................................................................38
2.11.5 Dye structure ..............................................................................................38
2.12 Fungal bioreactors for decolorization and degradation of dyes ............................38
2.13 Conclusion and future prospects ............................................................................39
References...............................................................................................................40
Further reading .......................................................................................................43
CHAPTER 3 Role of halophiles in xenobiotic bioremediation .............................. 45
Arunima Bhattacharjee, Aveepsa Sengupta, Sanchayita Basu,
Pragati Singh and Ashutosh Kumar
3.1 Xenobiotic compounds .............................................................................................45
3.2 Halophilesdan extremophle in action .....................................................................46
3.3 Metal bioremediation by halophiles .........................................................................49
3.4 Bioremediation of hydrocarbons including crude oils.............................................50
3.5 Bioremediation of crude oil......................................................................................51
3.6 Aliphatic hydrocarbon ..............................................................................................52
3.7 Monoaromatic hydrocarbon......................................................................................52
3.8 Polyaromatic hydrocarbon........................................................................................54
3.9 Conclusion ................................................................................................................55
References.................................................................................................................55
Further reading .........................................................................................................60
CHAPTER 4 Fungal diversity in the bioremediation of toxic effluents ................. 61
Mohita Chugh, Lakhan Kumar and Navneeta Bharadvaja
4.1 Introduction...............................................................................................................61
4.2 Methods of bioremediation.......................................................................................62
4.3 Steps of mycoremediation ........................................................................................63
4.3.1 Biosorption ..................................................................................................... 63
4.3.2 Bioaccumulation............................................................................................. 64
Contents vii
4.3.3 Biodegradation................................................................................................ 64
4.3.4 Bioconversion ................................................................................................. 65
4.4 Kinds of bioremediators ...........................................................................................65
4.4.1 White-rot fungi ............................................................................................... 65
4.4.2 Marine fungi ................................................................................................... 68
4.4.3 Extremophilic fungi........................................................................................ 69
4.4.4 Symbiotic fungi .............................................................................................. 69
4.4.5 Other kinds of mushrooms ............................................................................. 69
4.5 Factors affecting mycoremediation ..........................................................................70
4.5.1 Temperature effect on mycoremediation ....................................................... 70
4.5.2 pH effect on mycoremediation....................................................................... 70
4.5.3 Relative humidity effect on mycoremediation .............................................. 71
4.5.4 Effect of oxygen, light, trace elements, and aeration on
mycoremediation ............................................................................................ 71
4.6 Different types of targeted contaminants .................................................................71
4.6.1 Volatile organic compounds ........................................................................... 71
4.6.2 Synthetic dyes................................................................................................. 73
4.6.3 Heavy metals .................................................................................................. 74
4.6.4 Toxic compounds and municipal solid wastes .............................................. 74
4.7 Enzymes involved in bioremediation .......................................................................75
4.7.1 Peroxidase....................................................................................................... 76
4.7.2 Laccase ........................................................................................................... 77
4.7.3 Cytochrome P450 ........................................................................................... 77
4.8 Recent advancements in fungal bioremediation ......................................................78
4.9 Conclusion and future perspective ...........................................................................79
References.................................................................................................................80
Further reading .........................................................................................................88
CHAPTER 5 Current advances in microbial bioremediation of surface
and ground water contaminated by hydrocarbon.............................. 89
Micheal Bukola Alao and Elijah Adegoke Adebayo
5.1 Introduction...............................................................................................................89
5.1.1 Hydrocarbon (petroleum) contamination of surface water and
ground water................................................................................................... 90
5.1.2 Management of hydrocarbon contaminated water ........................................ 92
5.2 Microbial bioremediation of hydrocarbon (petroleum) contaminated water ..........94
5.2.1 Advantages of microbial bioremediation technique ...................................... 96
5.2.2 Current advances in microbial bioremediation technique ............................. 97
5.2.3 Mechanisms of bioremediating activity by microorganisms....................... 102
5.2.4 Current advances in application of microbial bioremediation .................... 105
5.3 Challenges and recommendations ..........................................................................109
5.3.1 Challenges .................................................................................................... 109
viii Contents
5.3.2 Recommendations ........................................................................................ 111

5.4 Conclusions and future prospects...........................................................................111
References...............................................................................................................112
Further reading .......................................................................................................116
CHAPTER 6 Microbial remediation of petroleum hydrocarbons in liquid
wastes ............................................................................................... 117
Rahul Saha and Soma Nag
6.1 Introduction...........................................................................................................117
6.2 Classification and composition of petroleum oil .................................................118
6.2.1 Very light oils .............................................................................................118
6.2.2 Light oils.....................................................................................................119
6.2.3 Medium oils................................................................................................119
6.2.4 Heavy fuel oils ...........................................................................................119
6.3 Impact of petroleum hydrocarbons on environment............................................119
6.4 Remediation techniques........................................................................................119
6.4.1 Physical techniques ....................................................................................120
6.4.2 Chemical techniques ..................................................................................120
6.4.3 Bioremediation techniques .........................................................................120
6.4.4 Phytoremediation........................................................................................120
6.5 Microbial degradation petroleum oil....................................................................121
6.6 Sources of microbes .............................................................................................122
6.7 Role of enzymes ...................................................................................................123
6.8 Role of biosurfactants in degrading hydrocarbon................................................124
6.9 Factors affecting the biodegradation of petroleum hydrocarbon.........................125
6.10 Advantages of microbial remediation technique .................................................125
6.11 Conclusion and future prospects ..........................................................................125
References.............................................................................................................126
Further reading .....................................................................................................129
CHAPTER 7 Microbial remediation of metals by marine bacteria ..................... 131
Sougata Ghosh, Joorie Bhattacharya, Rahul Nitnavare and
Thomas J. Webster
7.1 Introduction.............................................................................................................131
7.2 Heavy metal toxicity...............................................................................................132
7.3 Removal of toxic metals by marine microbes .......................................................134
7.3.1 Arsenic........................................................................................................ 134
7.3.2 Cadmium .................................................................................................... 134
7.3.3 Chromium ................................................................................................... 138
7.3.4 Cobalt.......................................................................................................... 138
7.3.5 Copper ........................................................................................................ 139
7.3.6 Iron.............................................................................................................. 141
Contents ix
7.3.7 Lead ............................................................................................................ 142

7.3.8 Manganese .................................................................................................. 145
7.3.9 Mercury ...................................................................................................... 146
7.3.10 Nickel.......................................................................................................... 148
7.3.11 Selenium ..................................................................................................... 149
7.3.12 Vanadium .................................................................................................... 151
7.3.13 Zinc ............................................................................................................. 152
7.4 Conclusions and future perspectives ......................................................................153
Acknowledgments...................................................................................................154
References...............................................................................................................154
Further reading .......................................................................................................158
CHAPTER 8 Microbial degradation of dye-containing wastewater .................... 159
Kumari Guddi and Angana Sarkar
8.1 Introduction...........................................................................................................159
8.2 Dye........................................................................................................................160
8.3 Classification of dyes............................................................................................160
8.4 Sources of hazardous dyes in the environment....................................................161
8.5 Mode of toxicity of dye in human health ............................................................164
8.6 Microbial interaction with dyes ...........................................................................165
8.7 Bacterial interaction with dye ..............................................................................166
8.8 Fungal interaction with dye..................................................................................166
8.9 Bioremediation strategies of dyes ........................................................................167
8.10 Bacterial degradation of dyes...............................................................................167
8.11 Fungal degradation of dyes ..................................................................................170
8.12 Hurdles on dye bioremediation ............................................................................171
8.13 Future prospects....................................................................................................172
References.............................................................................................................172
Further reading .....................................................................................................175
CHAPTER 9 Microbial bioremediation of heavy metals by Marine bacteria ..... 177
Raja Husain, Nitin Vikram, Garima Yadav, Deepak Kumar,
Sonika Pandey, Monika Patel, N.A. Khan and Touseef Hussain
9.1 Introduction...........................................................................................................177
9.2 Types of marine bacteria involved in metal bioremediation ...............................179
9.2.1 Diversity of marine bacteria.......................................................................180
9.3 Mechanism of microbial bioremediation of metals .............................................181
9.3.1 Remediation and metals .............................................................................181
9.4 Microbial bioremediation and its mechanism......................................................182
9.5 Microbial remediation of heavy metals from the soil .........................................183
9.6 Microbial bioremediation of metals from water..................................................184
9.7 Advantages and disadvantages of microbial bioremediation ..............................185
x Contents
9.8 Bioleaching (biomining).......................................................................................186

9.9 Phytoremediation ..................................................................................................187
9.10 Plant microbial bioremediation ............................................................................188
9.11 Heavy metal removal by fungi.............................................................................189
9.12 Metal uptake by fungi ..........................................................................................191
9.13 Heavy metal removal by Algae............................................................................193
9.14 Factors affecting heavy metal bioremediation by algae ......................................194
9.15 Factors affecting the microbial bioremediation of heavy metals ........................195
9.15.1 Physiochemical factors affecting bioremediation....................................196
9.15.2 Biological factors affecting bioremediation processes ............................196
9.15.3 Climate change and bioremediation processes........................................196
9.16 Conclusion and Future prospects .........................................................................196
Acknowledgment ..................................................................................................197
References.............................................................................................................197
Further reading .....................................................................................................203
CHAPTER 10 Role of microbes in biodegradation of cyanide and its metal
complexes ....................................................................................... 205
Vandita Anand and Anjana Pandey
10.1 Introduction...........................................................................................................205
10.2 Cyanide .................................................................................................................206
10.2.1 Chemistry of cyanide and their various forms ........................................206
10.2.2 Sources of cyanide ...................................................................................208
10.2.3 Cyanide discharge from industries and pollution ....................................209
10.2.4 Toxicological effects of cyanide ..............................................................211
10.2.5 Effluent disposal standards of cyanide.....................................................213
10.3 Factors responsible for the biodegradation of cyanides ......................................213
10.3.1 Concentration of cyanides........................................................................214
10.3.2 Availability of nutrients............................................................................214
10.3.3 Availability of oxygen ..............................................................................214
10.3.4 Presence of additional pollutants .............................................................215
10.3.5 Temperature and pH .................................................................................215
10.4 Microbial metabolism...........................................................................................215
10.4.1 Hydrolytic pathway ..................................................................................216
10.4.2 Oxidative pathway....................................................................................217
10.4.3 Reductive pathway ...................................................................................218
10.4.4 Substitution/transfer pathway...................................................................219
10.4.5 Synthases pathways ..................................................................................219
10.5 Advances in cyanide biodegradation technologies ..............................................220
10.6 Conclusion and future prospects ..........................................................................221
References.............................................................................................................222
Further reading .....................................................................................................224
Contents xi
CHAPTER 11 Microbial-mediated explosives removal and its impact

on TNT, RDX, and HMX ................................................................... 225
Komal Agrawal and Pradeep Verma
11.1 Introduction...........................................................................................................225
11.2 Classification of explosives ..................................................................................226
11.3 The problem with explosives ...............................................................................227
11.4 Impact of explosives on the environment ............................................................227
11.5 An overview of the environmental fate of explosives .........................................228
11.6 Bioremediation of explosives ...............................................................................229
11.6.1 TNT ..........................................................................................................229
11.6.2 RDX..........................................................................................................231
11.6.3 HMX .........................................................................................................233
11.7 Limitations, future prospects, and conclusion .....................................................234
References.............................................................................................................235
Further reading .....................................................................................................241
CHAPTER 12 Advancement in microbial bioremediation.................................... 243
Dixita Chettri, Bhaskar Sharma and Anil Kumar Verma
12.1 Introduction...........................................................................................................243
12.2 Xenobiotics and role of microbes ........................................................................244
12.2.1 Biodegradation..........................................................................................245
12.2.2 Biotransformation.....................................................................................245
12.2.3 Cometabolism...........................................................................................245
12.3 Parameters for biodegradation..............................................................................245
12.3.1 Microbial structure ...................................................................................246
12.3.2 Environmental factors ..............................................................................247
12.3.3 Physiochemical characteristic of the contaminant ..................................247
12.4 Approaches to bioremediation..............................................................................248
12.4.1 Cell-based bioremediation........................................................................248
12.4.2 Enzyme-based bioremediation .................................................................250
12.5 Improving the process of biodegradation.............................................................251
12.5.1 Genetic engineering..................................................................................251
12.5.2 Biosurfactants ...........................................................................................254
12.5.3 Chemotaxis and transport.........................................................................254
12.6 Omics approaches in the microbial bioremediation ............................................255
12.7 Conclusion and future prospective .......................................................................258
Acknowledgment ..................................................................................................258
References.............................................................................................................259
Further reading .....................................................................................................262
xii Contents
CHAPTER 13 Counterbalancing common explosive pollutants (TNT, RDX,

and HMX) in the environment by microbial degradation.............. 263
Nikita Chakraborty, Pakiza Begum and Bhisma Kumar Patel
13.1 Introduction...........................................................................................................263
13.1.1 Biological remediation .............................................................................264
13.1.2 Explosives.................................................................................................265
13.2 TNT.......................................................................................................................266
13.2.1 Chemistry and properties of TNT............................................................266
13.2.2 Worldwide deposits of TNT.....................................................................267
13.2.3 Toxicity of TNT .......................................................................................269
13.2.4 Microbial degradation of TNT.................................................................270
13.3 RDX and HMX.....................................................................................................286
13.3.1 Chemistry and properties of RDX and HMX..........................................286
13.3.2 Toxicity of RDX and HMX .....................................................................287
13.3.3 Biodegradation of RDX and HMX ..........................................................288
13.4 Conclusion ............................................................................................................301
13.5 Future research .....................................................................................................303
References.............................................................................................................304
CHAPTER 14 Enzyme-based biodegradation of toxic environmental
pollutants ........................................................................................ 311
Nahid Masood Siddiqui and Praveen Dahiya
14.1 Introduction...........................................................................................................311
14.2 Structure of enzymes ............................................................................................313
14.3 Mode of action of enzymes ..................................................................................314
14.3.1 Fisher template model (lock and key model) ..........................................314
14.3.2 Induced fit model......................................................................................314
14.4 Classification and nomenclature...........................................................................315
14.4.1 Oxidoreductase .........................................................................................315
14.4.2 Transferase................................................................................................316
14.4.3 Hydrolases ................................................................................................317
14.4.4 Lyases .......................................................................................................318
14.4.5 Isomerases ................................................................................................318
14.4.6 Ligases ......................................................................................................319
14.5 Industrial pollution and their impact on the ecosystem.......................................320
14.5.1 Water pollution .......................................................................................320
14.5.2 Air pollution ...........................................................................................321
14.5.3 Soil pollution ..........................................................................................322
14.5.4 Effects on plants .....................................................................................323
14.5.5 Effect on wildlife....................................................................................323
Contents xiii
14.5.6 Enzymes-based biodegradation of various industrial pollutants ...........324

14.5.7 Enzyme oxidoreductases ........................................................................324
14.5.8 Enzyme peroxidases ...............................................................................325
14.5.9 Enzyme laccases.....................................................................................327
14.5.10 Enzyme hydrolases.................................................................................327
14.5.11 Limitations and recent advances ............................................................328
14.6 Conclusion ............................................................................................................330
References.............................................................................................................330
Further reading .....................................................................................................333
CHAPTER 15 Cyanoremediation: a clean and green approach toward the
sustainable environment ................................................................ 335
Subhasish Dutta, Debojit Ghosh, Arunima Lahiri,
Sourish Chakraborty and Soumya Pandit
15.1 Introduction...........................................................................................................335
15.2 Cyanobacteria at a glance.....................................................................................338
15.2.1 Microbiology of cyanobacteria ................................................................338
15.3 Cyanobacteria as potential food, feed, and bioenergy source .............................338
15.4 Cyanoremediationda clean and green technology toward sustainable
future .....................................................................................................................339
15.4.1 Characteristics of cyanobacteria ..............................................................340
15.4.2 Different cyanoremediation approaches ..................................................340
15.5 Conclusion ............................................................................................................349
References.............................................................................................................350
Further reading .....................................................................................................354
CHAPTER 16 Enzymatic bioremediation: current status, challenges, future
prospects, and applications........................................................... 355
Luciana Porto de Souza Vandenberghe, Nelson Libardi Junior,
Kim Kley Valladares-Diestra, Susan Grace Karp,
Joyce Gueiros Wanderley Siqueira, Cristine Rodrigues and
Carlos Ricardo Soccol
16.1 Introduction...........................................................................................................355
16.2 Bioremediationdaspects and techniques.............................................................355
16.3 Enzymes for bioremediation ................................................................................358
16.3.1 Oxidoreductases........................................................................................358
16.3.2 Hydrolases ................................................................................................362
16.4 Modified enzymes for bioremediationdgenetic engineering/enzyme
engineering............................................................................................................367
16.5 Enzyme immobilization........................................................................................369
16.6 Advances on an innovation on enzymatic bioremediation ..................................371
xiv Contents
16.6.1 Nanozymes ...............................................................................................371

16.6.2 Classification of nanozymes.....................................................................372
16.6.3 Nanozymes applied in bioremediation processes ....................................373
16.6.4 Future prospects .......................................................................................374
References.............................................................................................................374
CHAPTER 17 An approach toward the biodegradation of PAHs by microbial
consortia ......................................................................................... 383
Subhasish Dutta and Sayan Laha
17.1 Introduction...........................................................................................................383
17.2 PAH-degrading pathways by different bacteria and fungi...................................385
17.2.1 Degradation mechanism of fungi.............................................................385
17.2.2 Nonligninolytic fungi mechanism............................................................389
17.2.3 PAHs degradation pathway by bacteria ...................................................392
17.3 Removal of PAHs from the soil by bacteria ........................................................395
17.3.1 Removal of some PAHs from soil ...........................................................395
17.3.2 Fenton’s reaction with viable cells ..........................................................395
17.3.3 Biostimulation and bioaugmentation .......................................................396
17.3.4 Remediation mechanism ..........................................................................396
17.3.5 Competition with microflora in contaminated soil..................................397
17.3.6 Competition ..............................................................................................398
17.4 Effect of incomplete combustion of PAHs on microbial and fungal
consortia................................................................................................................398
17.5 Proteomics and metabolomics in bioremediation of PAHs .................................399
17.6 Limiting factors of PAHs degradation .................................................................400
17.7 Remediation strategy for limiting factors ............................................................400
17.7.1 Choice of microorganism .........................................................................400
17.7.2 Bioavailability issue .................................................................................401
17.7.3 Other factors .............................................................................................402
17.8 Review on physicochemical treatments ...............................................................402
17.9 Conclusion and future aspects..............................................................................402
References.............................................................................................................403
Further reading .....................................................................................................406
CHAPTER 18 Bacterial- and fungal-mediated biodegradation of petroleum
hydrocarbons in soil....................................................................... 407
Vivek Kumar Gaur, Varsha Tripathi and Natesan Manickam
18.1 Introduction...........................................................................................................407
18.2 Composition of petroleum hydrocarbons.............................................................410
18.2.1 Aliphatics..................................................................................................411
18.2.2 Aromatics..................................................................................................412
Contents xv
18.2.3 Asphaltenes and waxes.............................................................................413

18.3 Impact of petroleum hydrocarbon on soil............................................................415
18.3.1 Soil fertility...............................................................................................415
18.3.2 Soil microflora ..........................................................................................415
18.4 Removal of petroleum hydrocarbon.....................................................................417
18.4.1 Bacterial biodegradation of petroleum hydrocarbon ...............................417
18.4.2 Fungal biodegradation of petroleum hydrocarbons .................................419
18.5 Conclusions...........................................................................................................421
Acknowledgments.................................................................................................421
References.............................................................................................................421
CHAPTER 19 Deep-marine bacteriadThe Frontier alternative for
heavy metals bioremediation ......................................................... 429
Neha Singh, Umesh Goutam and Mrinmoy Ghosh
19.1 Introduction...........................................................................................................429
19.2 Sources of heavy metals.......................................................................................431
19.2.1 Toxic effects of heavy metals on sea life ................................................432
19.3 Characteristics and diversity of deep-marine bacteria.........................................433
19.4 Deep-marine bacteria adaptation concerning changing environmental
conditions..............................................................................................................434
19.4.1 Factors affecting bioremediation process ................................................435
19.4.2 Mechanism of remediation processes ......................................................436
19.5 Heavy metal removal by deep-marine bacteria ...................................................439
19.6 Application of deep-marine bacteria in bioremediation ......................................440
19.7 Bioremediation enhancement via genetically modified deep-marine
bacteria..................................................................................................................441
19.7.1 Nickel........................................................................................................443
19.7.2 Cadmium ..................................................................................................443
19.7.3 Arsenic......................................................................................................443
19.7.4 Mercury ....................................................................................................444
19.8 Future prospective also pros and cons of using deep-marine bacteria................444
19.9 Conclusion ............................................................................................................446
References.............................................................................................................446
Further reading .....................................................................................................450
CHAPTER 20 Microbial biofilms for waste treatment and sustainable
development.................................................................................... 451
Arunima Bhattacharjee, Priya Khadgawat, Abhinav Suresh, Jolly Thomas,
Gayatri Brahmandam, Pragati Singh and Ashutosh Kumar
20.1 Introduction...........................................................................................................451
20.2 Biofilms in bioremediation...................................................................................452
xvi Contents
20.3 Oil bioremediation................................................................................................455

20.4 Textile wastewater treatment ................................................................................456
20.5 Removal of pharmaceuticals ................................................................................458
20.6 Bioremediation of persistent organic pollutants ..................................................458
20.7 Heavy metal remediation......................................................................................460
20.8 Limitations and future prospect ...........................................................................461
20.9 Conclusion ............................................................................................................461
References.............................................................................................................462
Further reading .....................................................................................................465
Index ................................................................................................................................................. 467

Contributors
Elijah Adegoke Adebayo
Department of Pure and Applied Biology, Microbiology Unit, Ladoke Akintola
University of Technology, Ogbomoso, Oyo, Nigeria
Komal Agrawal
Bioprocess and Bioenergy Laboratory, Department of Microbiology, Central
University of Rajasthan, Ajmer, Rajasthan, India
Micheal Bukola Alao
Department of Pure and Applied Biology, Microbiology Unit, Ladoke Akintola
University of Technology, Ogbomoso, Oyo, Nigeria
Vandita Anand
Department of Biotechnology, Motilal Nehru National Institute of Technology
(MNNIT) Allahabad, Prayagraj, Uttar Pradesh, India
Somdipta Bagchi
School of Infrastructure, Indian Institute of Technology Bhubaneswar,
Bhubaneswar, Odisha, India
Sanchayita Basu
Department of Microbiology, Tripura University (A Central University), Agartala,
Tripura, India
Pakiza Begum
Department of Chemistry, Gargaon College, Gargaon, Assam, India
Manaswini Behera
School of Infrastructure, Indian Institute of Technology Bhubaneswar,
Bhubaneswar, Odisha, India
Navneeta Bharadvaja
Department of Biotechnology, Delhi Technological University, Delhi, India
Arunima Bhattacharjee
Tripura, India
Joorie Bhattacharya
Genetic Gains, International Crops Research Institute for the Semi-Arid Tropics,
Hyderabad, Telangana, India
Gayatri Brahmandam
Sreenidhi Institute of Science and Technology, Hyderabad, Telangana, India
Nikita Chakraborty
Department of Chemistry, Indian Institute of Technology Guwahati, North
Guwahati, Assam, India
xvii
xviii Contributors
Sourish Chakraborty
Department of Biotechnology, Haldia Institute of Technology (HIT), ICARE
Complex, Haldia, West Bengal, India
Dixita Chettri
Department of Microbiology, Sikkim University, Gangtok, Sikkim, India
Mohita Chugh
Praveen Dahiya
Amity Institute of Biotechnology, Amity University Uttar Pradesh (AUUP), Noida,
India
Subhasish Dutta
Complex, Haldia, West Bengal, India; Chemical Engineering Division, Center of
Innovative and Applied Bioprocessing (CIAB), Mohali, Punjab, India
Vivek Kumar Gaur
Environmental Biotechnology Laboratory, Environmental Toxicology Group, CSIR-
Indian Institute of Toxicology Research, Lucknow, Uttar Pradesh, India; Amity
Institute of Biotechnology, Amity University Uttar Pradesh, Lucknow Campus,
Lucknow, Uttar Pradesh, India
Debojit Ghosh
Mrinmoy Ghosh
KIIT-Technology Business Incubator, Kalinga Institute of Industrial Technology
University, Bhubaneswar, Orissa, India
Sougata Ghosh
Department of Microbiology, School of Science, RK University, Rajkot, Gujarat,
India; Department of Chemical Engineering, Northeastern University, Boston,
MA, United States
Umesh Goutam
Department of Genetics Engineering and Molecular Biology, School of
Bioengineering and Biosciences, Lovely Professional University, Phagwara,
Punjab, India
Kumari Guddi
National Institute of Technology, Rourkela, Odisha, India
Contributors xix
Raja Husain
Department of Agriculture, Himalayan University, Itanagar, Arunachal Pradesh,
India; ICAR-Indian Institute of Pulses Research, Kanpur, Uttar Pradesh, India;
Department of Plant Molecular Biology and Genetic Engineering, Narendra Deva
University of Agriculture and Technology, Ayodhya, Uttar Pradesh, India
Touseef Hussain
Department of Botany, Aligarh Muslim University, Aligarh, Uttar Pradesh, India;
Division of Plant Pathology, ICAR-Indian Agricultural Research Institute, New
Delhi, Delhi, India
Nelson Libardi Junior
Departamento de Engenharia de Bioprocessos e Biotecnologia, Universidade
Federal do Paraná e UFPR, Curitiba, Parana, Brazil
Susan Grace Karp
Priya Khadgawat
Department of Genetics, University of Delhi, New Delhi, Delhi, India
N.A. Khan
Ashutosh Kumar
Tripura, India
Deepak Kumar
Department of Manufacturing & Development, Nextnode Bioscience Pvt. Ltd.,
Kadi, Gujarat, India; Department of Plant Molecular Biology and Genetic
Engineering, Narendra Deva University of Agriculture and Technology, Ayodhya,
Uttar Pradesh, India
Lakhan Kumar
Sayan Laha
Arunima Lahiri
xx Contributors
Natesan Manickam
Environmental Biotechnology Laboratory, Environmental Toxicology Group,
CSIR-Indian Institute of Toxicology Research, Lucknow, Uttar Pradesh, India
Soma Nag
Department of Chemical Engineering, National Institute of Technology Agartala,
Tripura, India
Rahul Nitnavare
Division of Plant and Crop Sciences, School of Biosciences, University of
Nottingham, Nottingham, United Kingdom; Department of Plant Sciences,
Rothamsted Research, Harpenden, United Kingdom
Anjana Pandey
Department of Biotechnology, Motilal Nehru National Institute of Technology
(MNNIT) Allahabad, Prayagraj, Uttar Pradesh, India
Sonika Pandey
ICAR-Indian Institute of Pulses Research, Kanpur, Uttar Pradesh, India
Soumya Pandit
Department of Life Sciences, School of Basic Sciences and Research, Sharda
University, Greater Noida, Uttar Pradesh, India
Bhisma Kumar Patel
Department of Chemistry, Indian Institute of Technology Guwahati, North
Guwahati, Assam, India
Monika Patel
CSIR-Central Drug Research Institute, Sector-10 Jankipuram Extension,
Lucknow, Uttar Pradesh, India
Bhumika Rajoria
Department of Pharmacology, University of Minnesota, Minneapolis, MN, United
States
Cristine Rodrigues
Arpita Roy
Department of Biotechnology, School of Engineering & Technology, Sharda
University, Greater Noida, Uttar Pradesh, India
Rahul Saha
Department of Chemical Engineering, National Institute of Technology Agartala,
Tripura, India
Angana Sarkar
National Institute of Technology, Rourkela, Odisha, India
Contributors xxi
Aveepsa Sengupta
Tripura, India
Bhaskar Sharma
School of Life and Environmental Sciences, Faculty of Science, Engineering, and
Built Environment, Deakin University, Geelong, Australia; TERI School of
Advanced Studies, New Delhi, Delhi, India
Nahid Masood Siddiqui
Amity Institute of Biotechnology, Amity University Uttar Pradesh (AUUP), Noida,
India
Neha Singh
Department of Genetics Engineering and Molecular Biology, School of
Bioengineering and Biosciences, Lovely Professional University, Phagwara,
Punjab, India
Pragati Singh
Tripura, India
Joyce Gueiros Wanderley Siqueira
Carlos Ricardo Soccol
Abhinav Suresh
Department of Biotechnology, Rajalakshmi Engineering College, Thandalam,
Tamil Nadu, India
Jolly Thomas
Center for Nanosciences and Molecular Medicine, Amrita Institute of Medical
Sciences and Research Center (AIMS), Kochi, Kerala, India
Varsha Tripathi
Environmental Biotechnology Laboratory, Environmental Toxicology Group,
CSIR-Indian Institute of Toxicology Research, Lucknow, Uttar Pradesh, India;
Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, Uttar
Pradesh, India
Kim Kley Valladares-Diestra
xxii Contributors
Luciana Porto de Souza Vandenberghe

Anil Kumar Verma
Department of Microbiology, Sikkim University, Gangtok, Sikkim, India
Pradeep Verma
Bioprocess and Bioenergy Laboratory, Department of Microbiology, Central
University of Rajasthan, Ajmer, Rajasthan, India
Nitin Vikram
Department of Agriculture, Zila Parishad Agriculture College, Banda, Uttar
Pradesh, India
Thomas J. Webster
Department of Chemical Engineering, Northeastern University, Boston, MA,
United States
Garima Yadav
ICAR-Indian Institute of Pulses Research, Kanpur, Uttar Pradesh, India;
CHAPTER
Microbial degradation of
xenobiotics in
bioelectrochemical
systems
1
Somdipta Bagchi, Manaswini Behera
School of Infrastructure, Indian Institute of Technology Bhubaneswar, Bhubaneswar, Odisha, India
1.1 Introduction to xenobiotics

The “xeno” in “xenobiotics” is a Greek work meaning guest, friend, or foreigner and
“bio” means life. Hence, xenobiotics are chemical compounds foreign to organisms
or environment. Xenobiotics are usually produced from human activities. However,
nature can also contribute to the production on these compounds. Endobiotic
(naturally occurring chemicals) when present in high concentration in the environ-
ment becomes xenobiotics. These are also produced by some organisms as a part of
their defense mechanism, for example, mycotoxin, bacterial and herbal toxin. These
have the ability to interact with environment by entering the food chain. Xenobiotics
can be chiefly classified into pesticides, polycyclic aromatic hydrocarbon (PAHs),
polychlorinated aromatics, solvents, surfactants, silicones, and plastics. Some of
the examples of nature’s contribution to formation of xenobiotics are seeping of
petroleum hydrocarbons through natural processes, production of volatile terpenes
and other halogenated compounds by plants, fungi, algae, bacteria, and other organ-
isms. They are also released into the environment from household, municipal,
agricultural, and industrial facilities. Although they are present in low concentra-
tions, these have become a concern because of their biorefractory nature. Total of
200,000 xenobiotic compounds have been identified so far. Xenobiotics have unnat-
ural structural features, making them nonbiodegradable or partially biodegradable.
The recalcitrance of the xenobiotic compounds can be attributed to
(i) nonrecognizable as substrate to degradable microorganisms, (ii) high stability
in water, (iii) insolubility in water, (iv) toxicity or toxic by-products generation as
a result of microbial degradation, (v) high molecular weight preventing entry to
microbial cells, and (vi) the unphysiological chemical bonds which block the attack
by microorganisms.
Human exposure to xenobiotics is unavoidable, as they are used as drugs,
antibiotics, dietary supplements like antioxidants. They are also used as food ingre-
dients (dyes, stabilizers, preservatives, emulsifier, salt compounds), cosmetics, and
personal care and household products (makeup, perfumes, soaps, bleach, sprays,
Development in Wastewater Treatment Research and Processes. https://doi.org/10.1016/B978-0-323-85839-7.00020-7 1

Copyright © 2022 Elsevier Inc. All rights reserved.
2 CHAPTER 1 Microbial degradation of xenobiotics
and cleaners) on a daily basis. The industrial, domestic, and agricultural wastes run
off into rivers, lakes, and groundwater and are consumed by human beings, animals,
and used for agricultural purposes. The mode of contamination is classified into
direct and indirect. Direct contamination is from water collected from polluted water
reserves, whereas indirect contamination is from consumption of vegetables,
cereals, and fruits raised using fertilizer sludge from sewage water treatment unit
or noncomposted human or animal dung. Consumption of meat, fish, shrimps, or
snails fed with or collected from polluted water bodies also leads to indirect
consumption. These xenobiotics presently do not have any regulatory standards in
most of the countries as they are usually present at an extremely low concentration
in the aquatic environment. The xenobiotics after consumption causes biotoxicity,
bioassimilation, genetic mutations and chronic diseases, etc., in all life forms.
Because of its biorefractory nature, xenobiotics are often emitted nondegraded in
conventional primary physical operations and biological secondary wastewater
treatment processes and appear in the effluents of wastewater treatment plants in
the absence of an efficient tertiary treatment unit. The various tertiary treatment
processes, such as membrane filtration, adsorption, advanced oxidation, etc., can
efficiently remove xenobiotics but they require huge amount of energy, economy,
maintenance, and generate toxic by-products and sludge, hence are not
recommended.
Bioelectrochemical system (BES) is a state-of-the-art technology, which recently
has gained a lot of attention in the light of degradation of xenobiotics. The BES are
devices that can convert chemical energy into electrical energy employing microor-
ganisms as catalysts or utilizes electrical energy to produce hydrogen, methane, or
many other compounds of interest from organic matter which are called microbial
fuel cell (MFC) and microbial electrolysis cell (MEC), respectively. A typical
double-chambered MFC consists of an anode and cathode chamber separated
most commonly by a proton exchange membrane (PEM). A current collector with
an external load connects the anode and cathode. The anode chamber of the MFC
is usually inoculated with anaerobic sludge. The substrate added in the anode
undergoes anaerobic digestion by exoelectrogens producing protons, electrons,
and CO2. The electrons released are transferred through mediator, conductive pilli,
or nanowires to the anode, which then flows to the cathode via the current collector.
The protons generated in the anodic chamber migrate to the cathodic chamber
crossing the PEM to maintain charge equilibrium in the system. Whereas in
MEC, electricity is supplied to the system to obtain hydrogen, methane, or any other
value-added products from the organic matter present in the wastewater. In micro-
bial desalination cell (MDC), a third chamber is introduced between the anode
and cathode chambers. This middle chamber is called a desalination chamber where
saline water is added. Anion exchange membrane (AEM) next to the anode and a
cation exchange membrane (CEM) next to the cathode are placed in both the sides
of the desalination chamber. When the bacteria degrade the organic matter at the
anode, transferring free electrons through the external circuit, a positively charged
medium in the anode is created along with creation of a negatively charged medium
1.2 Conventional processes of xenobiotics degradation 3
in the cathode chamber. This potential difference between anode and cathode cham-
ber drives the flow of anions (Cl) from the desalination chamber through the AEM
into the anode and the flow of cations (Naþ) through the CEM into the cathode
thereby resulting in the desalination of water in the desalination chamber.
The combination of oxidizing as well as reducing environment in BES and the
robust microbial consortia in the form of inoculum makes BES a suitable choice
for xenobiotics degradation in comparison to other conventional processes.
Furthermore, the generation of highly reactive species such as OOH, OH, etc.,
via a two-electron pathway and suitable modifications such as utilization of cathode
catalyst leads to increased production of reactive oxygen species improving the
degradation efficiency of xenobiotics (Chakraborty et al., 2020).
Hence, in this chapter, the three BES, namely MFC, MEC, and MDC, have been
discussed to understand the feasibility xenobiotics degradation and power
generation using these three technologies.
1.2 Conventional processes of xenobiotics degradation and

their drawbacks
Xenobiotics are released untreated or partially treated from the primary treatment
units and owing to their recalcitrance they can only be treated completely in
advanced processes. The different xenobiotic compounds such as pharmaceuticals
and personal care products, contrast media, plasticizers, food additives, wood
preservatives, laundry detergents, surfactants, disinfectants, flame retardants, pesti-
cides, natural and synthetic hormones, and a few disinfection by-products, etc., are
usually discharged by households and industries. Because of their recalcitrance,
these xenobiotic compounds or their metabolites end up in the environment, due
to which the awareness regarding the influence of these pollutants on environmental
water bodies are rising and their treatment is considered important. The presence of
the xenobiotics in aquatic bodies can have a lot of ecotoxicological effects and high
risks to human health due to its bioaccumulation and microbial drug resistance. It
was reported that the compounds bromoform, chloroform, diclofenac, caffeine,
ibuprofen, naproxen, methyl dihydro jasmonate, galaxolide, butylated hydroxyto-
luene, and butylated hydroxyanisole were found in irrigation water and later in
the plants. Hormones, which act as endocrine disruption agents, were reported to
affect the reproductive health and survival of several species of fishes. The following
conventional treatment processes have been till date used for the treatment of
xenobiotics in wastewater (Fetzner, 2002).
1.2.1 Adsorption
The process of adsorption has been used a lot recently to degrade xenobiotics
because of its advantages of simplicity in operation and designing, capability to
handle microlevel of pollutants, toxicity removal, low investment cost,
environmentally benign nature, and the probability of adsorbent reuse and regener-
ation. The exploration for new adsorbents has been intensified in recent years. The
various xenobiotic compounds being treated using adsorption are naproxen (NPX),
clofibric acid, bisphenol-A (BPA), tetracycline antibiotic, and diclofenac. Adsorp-
tion has also been used to treat xenobiotics from water and wastewater sludge, paper
mill wastewaters, sediments and soil, fragrance materials, pesticides, and human
pharmaceuticals from environment. The various adsorbent materials used till date
are zeolites, organic soils, organic carbon framework, metal organic framework,
activated carbon, biochar, activated hydrochars, carbon nanotubes, graphenes,
composites with activated carbon, mesoporous nanocomposite of polymer and
clay, mesoporous silica, tailored composites, nanocomposites of carbon nano-
tubes/nanofibers, and layered silicates. The process of transfer of the xenobiotic
onto the solid surface of the adsorbent is called adsorption. The characteristics of
adsorbent and adsorbate are unambiguous, and adsorption depends upon their
composition and chemical nature, respectively. Adsorption depends on the compo-
sition and chemical nature of the adsorbent and adsorbate and involves interactions
such as van der Waals forces of interaction, electrostatic attraction, pep interac-
tions, and other hydrophobic interactions (Lima, 2018).
1.2.2 Membrane processes

Membrane processes, including forward osmosis (FO), reverse osmosis (RO),
nanofiltration (NF), and ultrafiltration (UF), have been widely used in water and
wastewater treatment processes. In FO high-quality permeate is produced because
of its ability to operate under an osmotic driving force without requiring a hydraulic
pressure difference, whereas in RO, the separation of xenobiotics happens through
adsorption onto the membrane surfaces followed by dissolution into the membrane,
and subsequent transport of xenobiotics through the membrane matrix. In NF, the
retention of xenobiotics depends on the properties of the compounds and the chem-
istry of the solution. In UF membrane processes, the separation mechanisms of size/
steric exclusion, hydrophobic adsorption, and electrostatic repulsion have been
reported to be used to remove both organic and inorganic xenobiotics (Semião
and Schäfer, 2010).
1.2.3 Advanced oxidation processes

Advanced oxidation processes (AOPs) such as ozonation (O3/H2O2), photocatalysis
(UV/TiO2), ultraviolet (UV/H2O2), and Fenton/photo-Fenton, and ultrasonic (US)
treatment have been recognized as an advanced treatment process for the removal
of xenobiotics from water and wastewater. AOP treatment can happen either via
generation of hydroxyl radical (OH) or other oxidative radicals which react with
the xenobiotics to mineralize them into carbon dioxide, water, and inorganic ions.
Ozonation (O3) along with other ozone combinations such as (O3/H2O2), (O3/UV),
or (O3/H2O2/UV), with and without catalysts, has been extensively used to study
1.2 Conventional processes of xenobiotics degradation 5
removal of antibiotics from wastewater. Ozone at high pH increases the production

of OH which can efficiently remove antibiotics from wastewater. Ozone in
combination with hydrogen peroxide has been reported to remove a wide range of
pollutants in water and wastewater. The factors affecting the efficiency of ozoniza-
tion are pH, ozone dosage, and temperature. While ozonation can recover water up to
99%, it has the drawback of high energy demand and high cost to implement. Another
AOP, namely Fenton processes, was also been reported to be used for treatment of
antibiotics at laboratory and plant/pilot scale. During the process, reaction occurs
in the acidic medium (2.8e3.0) in presence of H2O2 and Fe2þ producing OH which
leads to oxidation of organic pollutants. Fenton processes are economic, safe, quick,
and environmentally friendly as iron is already present in plenty. The antibiotic
ofloxacin was reported to be removed 100% by Fenton process. Photo Fenton process
occurs in the presence of UV radiation or sunlight with the combination of H2O2 and
Fe2þ or Fe3þ to enhance the production of OH. Photo-fenton processes have been
reported to remove many pharmaceutical compounds (PhACs) with higher removal
efficiencies of 95%e100% except penicillin G. The combination of various AOPs
has recently gained attention for the removal of antibiotics in wastewater. The
integration of AOPs with other compounds or processes like hydrogen peroxide,
catalyst, solar, or UV enhances the removal efficiency of the process. It has been
reported that UV/H2O2 and ozonation method can degrade antibiotics up to 100%
except metronidazole (92%) and ciprofloxacin (93%). Similarly, UV/H2O2/Fe2þ
showed the degradation value up to 100% in antibiotics except clarithromycin
(93%). It has been observed from the previous studies that the industrial level
application on the removal of antibiotics is not much performed and further studies
are required to identify the intermediates or by-products being discharged into the
environment (Anjali and Santhakumar, 2019).
1.2.4 Biological treatment processes

Several different biological processes are available, with activated sludge systems
being the most commonly used technology to treat xenobiotics. The processes can
be either aerobic or anaerobic processes depending on the type of contaminant.
These biological processes are often coupled with other tertiary treatment processes.
Apart from activated sludge process, the other biological processes such as biolog-
ical filtration have been tested for the degradation of xenobiotics. The selection of
the nature of the process employed (aerobic or anaerobic) often depends on the ter-
minal electron-accepting conditions required. For example, benzotriazoles seem to
be better removed under aerobic conditions in comparison to anaerobic conditions
with varied response with different electron acceptors. Apart from electron accep-
tors, operating temperatures also play a huge role in degrading the xenobiotics, as
natural and synthetic estrogen and nonylphenol derivatives have been reported to
be degraded 60%e90% under mesophilic and thermophilic conditions using anaer-
obic processes. However, there is lack of knowledge regarding extraction methodol-
ogies of xenobiotics from biological processes. The different biological treatment
processes including conventional activated sludge, biological nutrient removal, and

membrane bioreactors have showed that only easily biodegradable ECs (e.g.,
caffeine, diclofenac, trimethoprim) can be removed, whereas low biodegradables
(e.g., sulpiride, metoprolol, bezafibrate) may not be eliminated at all by biological
processes. The lack of comparable results, relatively limited information on the
microorganisms involved in the degradation process, toxicity of the targeted
compounds on the microorganisms involved, biofouling, handling of biosolids pro-
duced, and influence of other compounds present in the wastewater effluents on the
overall xenobiotics removal efficiencies are some of the drawbacks of the conven-
tional biological processes (Rodriguez-Narvaez et al., 2017).
1.3 Xenobiotics degradation in bioelectrochemical

systems
1.3.1 Microbial fuel cell
Xenobiotic compounds can be either oxidized in anodic or reductively cleaved at the
cathodic chamber of the dual-chambered MFC depending on the nature of the com-
pound. Following section contains a review on degradation of some of most
commonly present xenobiotics in environment. Fig. 1.1 shows the schematic of an
MFC (Fig. 1.1).
1.3.1.1 Pharmaceuticals
PhACs are usually present in trace concentration in wastewater and can be removed
very efficiently in the electrogenic and combined oxidative/reductive environment
LOAD
e e
O2
Organics -
+
OH
H
+
H H2O
CO2
Anode Cathode
chamber CEM chamber
FIGURE 1.1
Schematic of microbial fuel cell.
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“Well, I’m glad you’ve got here. We’ve been having a picnic up at
the house. Julie’s been having the hysterics and MacDonald—you
never knew MacDonald, did you?”
Applegate listened politely. He had a curious feeling that Julie and
her hysterics were already very far away and unimportant to him, but
he did not wish to be so brutal as to show this.
“When did MacDonald return and where has he been?” he asked,
gravely.
“He got here yesterday. He says he had a shock or something in
that accident—anyhow, he just couldn’t remember anything, and
when he come to he didn’t know who he was, nor anything about
himself, and all his papers and clothes had been burnt, so there was
nothing to show anybody who he was. He could work, and he was all
right most ways. Says he was that way till about six months ago,
when a Frisco doctor got hold of him and did something to his head
that put him right. He has papers from the doctor to show it’s true.
His case attracted lots of attention out there. Of course he wrote to
Julie when he came to himself, but his letters went to our old
address and she never got them. So then he started East to see
about it. He says he’s got into a good business and is going to do
well.”
There was a long silence. Presently Hopson began again,
awkwardly:
“I don’t know how you feel about it, but I think Julie’d ought to go
back to him.”
Applegate’s heart began to beat in curious, irregular throbs; he
could feel the pulsing of the arteries in his neck and there was a
singing in his ears.
“Of course Julie agrees with you?” he said, thickly.
“Well, no; she don’t. That’s what she wanted me to talk to you
about. She can’t see it but one way. She says he died, or if he didn’t
it was the same thing to her, and she married you. She says nobody
can have two husbands, and it’s you who are hers. I told her the law
didn’t look at it that way, and she says then she must get a divorce
from MacDonald and remarry you. MacDonald says if she brings suit
on the ground of desertion he will fight it. He says he can prove it
ain’t been no wilful desertion. But probably he could be brought
round if he saw she wouldn’t go back to him anyhow. MacDonald
wouldn’t be spiteful. But he was pretty fond of Julie.”
Applegate had stopped suddenly in the middle of Hopson’s
speech. Now he went forward rapidly, but he made no answer.
Hopson scrutinized his face a moment before he continued:
“Julie says you won’t be spiteful either. She says maybe she was a
little hasty in what she said just before she came up here. But you
know Julie’s way.”
“Yes,” said Applegate, “I know Julie’s way.”
Hopson drew a breath of relief. He had at least discharged himself
of his intercessory mission.
“I tell Julie she’d better put up with it and go with MacDonald. The
life would be more the sort of thing she likes. But her head’s set and
she won’t hear to anything Henriette or I say. You see, that’s what
Julie holds by, what she thinks is respectable. And it’s about all she
does hold by.” He hesitated, groping blindly about in his
consciousness for words to express his feeling that this passionate,
reckless nature was only anchored to the better things of life by her
fervent belief in the righteousness of the established social order.
“Julie thinks everything of being respectable,” he concluded,
lamely.
“Is it much farther to your house?” asked Applegate, dully.
“Right here,” answered Hopson, pulling his key from his pocket.
They entered a crude little parlor whose carpet was too gaudy, and
whose plush furniture was too obviously purchased at a bargain, but
its air was none the less heavy with tragedy. A single gas-jet
flickered in the centre of the room. On one side a great, broad-
shouldered fellow sat doggedly with his elbows on his knees and his
face buried in his hands. There was resistance in every line of his
figure. On the sofa opposite was Julie in her crimson dress. As she
lifted her face eagerly, Applegate noticed traces of tears upon it. Mrs.
Hopson, who had been moving about the room aimlessly, a pale and
ineffective figure between these two vivid personalities, came to a
standstill and looked at Applegate breathlessly. For a moment no
one spoke. Then Julie, baffled by the eyes she could not read,
sprang to her feet and stretched out her hands with a vehement
gesture.
“John Applegate, you’ll put me right! You will. I know you will. I
can’t go back to him! How can I?” Her hungry eyes scrutinized his
still, inexpressive face.
“John, you aren’t going to turn me off?” Her voice had a despairing
passion in it. “You won’t refuse to marry me if I get the divorce?
Good God! You can’t be such a devil. John! oh, John!”
Applegate sat down and looked at her apathetically. He was not
used to being called a devil. Somehow it seemed to him the term
was misapplied.
“Don’t take on so, Julie,” he said, quietly. The room seemed to
whirl around him, and he added, with a palpable effort:
“I’ll think it over and try to do what is best for both of us.”
At that MacDonald lifted his sullen face from his hands for the first
time and glanced across at the other man with blood-shot eyes.
Then he rose slowly, his great bulk seeming to fill the room, and
walking over to Applegate’s chair stood in front of it looking down at
him. His scrutiny was long. Once Applegate looked up and met his
eyes, but he was too tired to bear their fierce light and dropped his
own lids wearily.
MacDonald turned from him contemptuously and faced his wife,
who averted her head.
“Look at me, Julie!” he cried, appealingly. “I am better worth it than
he is. Good Lord! I don’t see what you see in him. He’s so tame! Let
him go about his business. He’s nobody. He don’t want you. Come
along with me and we’ll lead a life! You shall cut a dash out there. I
can make money hand over fist. It’s the place for you. Come on!”
For a moment Julie’s eyes glittered. The words allured her, but her
old gods prevailed. She threw out her arms as if to ward off his
proposal.
“No, no,” she said, shrilly. “I cannot make it seem right. You were
dead to me, and I married him. One does not go back to the dead. If
I am your wife, what am I to him? It puts me in the wrong these two
years. I cannot have it so, I tell you. I cannot have it so!”
Applegate felt faint and sick. Rising, he groped for the door. “I
must have air,” he said to Hopson, confusedly. “I will come back in a
minute.”
Once outside, the cool November night refreshed him. He dropped
down upon the doorstep and threw back his head, drinking in long
breaths as he looked up at the mocking stars.
When he found at last the courage to ask himself what he was
going to do, the answer was not ready. The decision lay entirely in
his hands. He might still be free if he said the word; and as he
thought of this he trembled. He had always tried to be what his
neighbors called a straight man, and he wanted to be straight in this
also. But where, in such a hideous tangle, was the real morality to be
found? Surely not in acceding to Julie’s demands! What claim had
she upon the home whose simple traditions of peace and happiness
she had trampled rudely under foot? Was it not a poor, cheap
convention of righteousness which demanded he should take such a
woman back to embitter the rest of his days and warp his children’s
lives? He rebelled hotly at the thought. That it was Julie’s view of the
ethical requirement of her position made it all the more improbable
that it was really right. Surely his duty was to his children first, and as
for Julie, let her reap the reward of her own temperament. The Lord
God Himself could not say that this was unjust, for it is so that He
deals with the souls of men.
It seemed to him that he had decided, but as he rose and turned to
the door a new thought stabbed him so sharply that he dropped his
lifted hand with a groan.
Where had been that sense of duty to his children, just now so
imperative, in the days when he had yielded to Julie’s charm against
his better judgment? Had duty ever prevailed against inclination with
him? Was it prevailing now?
High over all the turmoil and desperation of his thoughts shone out
a fresh perception that mocked him as the winter stars had mocked.
For that hour at least, the crucial one of his decision, he felt assured
that in the relation of man and woman to each other lies the supreme
ethical test of each, and in that relation there is no room for
selfishness. It might be, indeed, that he owed Julie nothing, but
might it not also be that the consideration he owed all womankind
could only be paid through this woman he had called his wife? This
was an ideal with which he had never had to reckon.
He turned and sat him down again to fight the fight with a chill
suspicion in his heart of what the end would be.
Being a plain man he had only plain words in which to phrase his
decision when at last he came to it.
“I chose her and I’ll bear the consequences of my choice,” he said,
“but I’ll bear them by myself. His aunt will be glad to take Teddy, and
Dora is old enough to go away to school.” Then he opened the door.
Hopson and his wife had left the little parlor. Julie on the sofa had
fallen into the deep sleep of exhaustion. MacDonald still sat there,
with his head in his hands, and to him Applegate turned. At the
sound of his step the man lifted his massive head and shook it
impatiently.
“Well?” he demanded.
“The fact is, Mr. MacDonald, Julie and I don’t get along very well
together, but I don’t know as that is any reason why I should force
her to do anything that don’t seem right to her. She thinks it would be
more”—he hesitated for a word—“more nearly right to get a divorce
from you and remarry me. As I see it now, it’s for her to say what she
wants, and for you and me to do it.”
MacDonald looked at him piercingly.
“You know you’d be glad of the chance to get rid of her!” he
exclaimed, excitedly. “In Heaven’s name, then, why don’t you make
her come to me? You know I suit her best. You know she’s my sort,
not yours. She’s as uncomfortable with you as you with her, and
she’d soon get over the feeling she has against me. Man! There’s no
use in it! Why can’t you give my own to me?”
“I can’t say I don’t agree with you,” said Applegate, and the words
seem to ooze painfully from his white lips, “but she thinks she’d
rather not, and—it’s for her to say.”
A CONSUMING FIRE
He is a man who has failed in this life, and says he has no chance
of success in another; but out of the fragments of his failures he has
pieced together for himself a fabric of existence more satisfying than
most of us make of our successes. It is a kind of triumph to look as
he does, to have his manner, and to preserve his attitude toward
advancing years—those dreaded years which he faces with pale but
smiling lips.
If you would see my friend Hayden, commonly called by his friends
the connoisseur, figure to yourself a tall gentleman of sixty-five, very
erect still and graceful, gray-headed and gray-bearded, with fine gray
eyes that have the storm-tossed look of clouds on a windy March
day, and a bearing that somehow impresses you with an idea of the
gracious and pathetic dignity of his lonely age.
I myself am a quiet young man, with but one gift—I am a finished
and artistic listener. It is this talent of mine which wins for me a
degree of Hayden’s esteem and a place at his table when he has a
new story to tell. His connoisseurship extends to everything of
human interest, and his stories are often of the best.
The last time that I had the honor of dining with him, there was
present, besides the host and myself, only his close friend, that
vigorous and successful man, Dr. Richard Langworthy, the eminent
alienist and specialist in nervous diseases. The connoisseur
evidently had something to relate, but he refused to give it to us until
the pretty dinner was over. Hayden’s dinners are always pretty, and
he has ideals in the matter of china, glass, and napery which it would
require a woman to appreciate. It is one of his accomplishments that
he manages to live like a gentleman and entertain his friends on an
income which most people find quite inadequate for the purpose.
After dinner we took coffee and cigars in the library.
On the table, full in the mellow light of the great lamp (Hayden has
a distaste for gas), was a bit of white plush on which two large opals
were lying. One was an intensely brilliant globe of broken gleaming
lights, in which the red flame burned strongest and most steadily; the
other was as large, but paler. You would have said that the prisoned
heart of fire within it had ceased to throb against the outer rim of ice.
Langworthy, who is wise in gems, bent over them with an
exclamation of delight.
“Fine stones,” he said; “where did you pick them up, Hayden?”
Hayden, standing with one hand on Langworthy’s shoulder, smiled
down on the opals with a singular expression. It was as if he looked
into beloved eyes for an answering smile.
“They came into my possession in a singular way, very singular. It
interested me immensely, and I want to tell you about it, and ask
your advice on something connected with it. I am afraid you people
will hardly care for the story as much as I do. It’s—it’s a little too
rococo and sublimated to please you, Langworthy. But here it is:
“When I was in the West last summer, I spent some time in a city
on the Pacific slope which has more pawnbrokers’ shops and that
sort of thing in full sight on the prominent streets than any other town
of the same size and respectability that I have ever seen. One day,
when I had been looking in the bazaars for something a little out of
the regular line in Chinese curios and didn’t find it, it occurred to me
that in such a cosmopolitan town there might possibly be some
interesting things in the pawn-shops, so I went into one to look. It
was a common, dingy place, kept by a common, dingy man with
shrewd eyes and a coarse mouth. Talking to him across the counter
was a man of another type. Distinction in good clothes, you know,
one is never sure of. It may be only that a man’s tailor is
distinguished. But distinction in indifferent garments is distinction
indeed, and there before me I saw it. A young, slight, carelessly
dressed man, his bearing was attractive and noteworthy beyond
anything I can express. His appearance was perhaps a little too
unusual, for the contrast between his soft, straw-colored hair and
wine-brown eyes was such a striking one that it attracted attention
from the real beauty of his face. The delicacy of a cameo is rough,”
added the connoisseur, parenthetically, “compared to the delicacy of
outline and feature in a face that thought, and perhaps suffering,
have worn away, but this is one of the distinctive attractions of the
old. You do not look for it in young faces such as this.
“On the desk between the two men lay a fine opal—this one,” said
Hayden, touching the more brilliant of the two stones. “The younger
man was talking eagerly, fingering the gem lightly as he spoke. I
inferred that he was offering to sell or pawn it.
“The proprietor, seeing that I waited, apparently cut the young man
short. He started, and caught up the stone. ‘I’ll give you—’ I heard
the other say, but the young man shook his head, and departed
abruptly. I found nothing that I wanted in the place, and soon passed
out.
“In front of a shop-window a little farther down the street stood the
other man, looking in listlessly with eyes that evidently saw nothing.
As I came by he turned and looked into my face. His eyes fixed me
as the Ancient Mariner’s did the Wedding Guest. It was an appealing
yet commanding look, and I—I felt constrained to stop. I couldn’t
help it, you know. Even at my age one is not beyond feeling the force
of an imperious attraction, and when you are past sixty you ought to
be thankful on your knees for any emotion that is imperative in its
nature. So I stopped beside him. I said: ‘It is a fine stone you were
showing that man. I have a great fondness for opals. May I ask if you
were offering it for sale?’
“He continued to look at me, inspecting me calmly, with a
fastidious expression. Upon my word, I felt singularly honored when,
at the end of a minute or two, he said: ‘I should like to show it to you.
If you will come to my room with me, you may see that, and another;’
and he turned and led the way, I following quite humbly and gladly,
though surprised at myself.
“The room, somewhat to my astonishment, proved to be a large
apartment—a front room high up in one of the best hotels. There
were a good many things lying about which obviously were not hotel
furnishings, and the walls, the bed, and even the floor were covered
with a litter of water-color sketches. Those that I could see were
admirable, being chiefly impressions of delicate and fleeting
atmospheric effects.
“I took the chair he offered. He stood, still looking at me,
apparently not in haste to show me the opals. I looked about the
room.
“‘You are an artist?’ I said.
“‘Oh, I used to be, when I was alive,’ he answered, drearily. ‘I am
nothing now.’ And then turning away he fetched a little leather case,
and placed the two opals on the table before me.
“‘This is the one I have always worn,’ he said, indicating the more
brilliant. ‘That chillier one I gave once to the woman whom I loved. It
was more vivid then. They are strange stones—strange stones.’
“He said nothing more, and I sat in perfect silence, only dreading
that he should not speak again. I am not making you understand
how he impressed me. In the delicate, hopeless patience of his face,
in the refined, uninsistent accents of his voice, there was somehow
struck a note of self-abnegation, of aloofness from the world,
pathetic in any one so young.
“I am old. There is little in life that I care for. My interests are
largely affected. Wine does not warm me now, and beauty seems no
longer beautiful; but I thank Heaven I am not beyond the reach of a
penetrating human personality. I have at least the ordinary instincts
for convention in social matters, but I assure you it seemed not in the
least strange to me that I should be sitting in the private apartment of
a man whom I had met only half an hour before, and then in a
pawnbroker’s shop, listening eagerly for his account of matters
wholly personal to himself. It struck me as the most natural and
charming thing in the world. It was just such chance passing
intercourse as I expect to hold with wandering spirits on the green
hills of paradise.
“It was some time before he spoke again.
“‘I saw her first,’ he said, looking at the paler opal, as if it was of
that he spoke, ‘on the street in Florence. It was a day in April, and
the air was liquid gold. She was looking at the Campanile, as if she
were akin to it. It was the friendly grace of one lily looking at another.
Later, I met her as one meets other people, and was presented to
her. And after that the days went fast. I think she was the sweetest
woman God ever made. I sometimes wonder how He came to think
of her. Whatever you may have missed in life,’ he said, lifting calm
eyes to mine, and smiling a little, ‘you whose aspect is so sweet,
decorous, and depressing, whose griefs, if you have griefs, are the
subtle sorrows of the old and unimpassioned’—I remember his
phrases literally. I thought them striking and descriptive,” confessed
Hayden—“‘I hope you have not missed that last touch of exaltation
which I knew then. It is the most exquisite thing in life. The Fates
must hate those from whose lips they keep that cup.’ He mused
awhile and added, ‘There is only one real want in life, and that is
comradeship—comradeship with the divine, and that we call religion;
with the human, and that we call love.’
“‘Your definitions are literature,’ I ventured to suggest, ‘but they are
not fact. Believe me, neither love nor religion is exactly what you call
it. And there are other things almost as good in life, as surely you
must know. There is art, and there is work which is work only, and
yet is good.’
“‘You speak from your own experience?’ he said, simply.
“It was a home thrust. I did not, and I knew I did not. I am sixty-five
years old, and I have never known just that complete satisfaction
which I believe arises from the perfect performance of distasteful
work. I said so. He smiled.
“‘I knew it when I set my eyes upon you, and I knew you would
listen to me and my vaporing. Your sympathy with me is what you
feel toward all forms of weakness, and in the last analysis it is self-
sympathy. You are beautiful, not strong,’ he added, with an air of
finality, ‘and I—I am like you. If I had been a strong man.... Christ!’
“I enjoyed this singular analysis of myself, but I wanted something
else.
“‘You were telling me of the opals,’ I suggested.
“‘The opals, yes. Opals always made me happy, you know. While I
wore one, I felt a friend was near. My father found these in Hungary,
and sent them to me—two perfect jewels. He said they were the twin
halves of a single stone. I believe it to be true. Their mutual relation
is an odd one. One has paled as the other brightened. You see them
now. When they were both mine, they were of almost equal
brilliancy. This,’ touching the paler, ‘is the one I gave to her. You see
the difference in them now. Hers began to pale before she had worn
it a month. I do not try to explain it, not even on the ground of the old
superstition. It was not her fault that they made her send it back to
me. But the fact remains; her opal is fading slowly; mine is burning to
a deeper red. Some day hers will be frozen quite, while mine—mine
—’ his voice wavered and fell on silence, as the flame of a candle
fighting against the wind flickers and goes out.
“I waited many minutes for him to speak again, but the silence was
unbroken. At last I rose. ‘Surely you did not mean to part with either
stone?’ I said.
“He looked up as if from a dream. ‘Part with them? Why should I
sell my soul? I would not part with them if I were starving. I had a
minute’s temptation, but that is past now.’ Then, with a change of
manner, ‘You are going?’ He rose with a gesture that I felt then and
still feel as a benediction. ‘Good-by. I wish for your own sake that
you had not been so like my poor self that I knew you for a friend.’
“We had exchanged cards, but I did not see or hear of him again.
Last week these stones came to me, sent by some one here in New
York of his own name—his executor. He is dead, and left me these.
“It is here that I want your counsel. These stones do not belong to
me, you know. It is true that we are like, as like as blue and violet.
But there is that woman somewhere—I don’t know where; and I
know no more of their story than he told me. I have not cared to be
curious regarding it or him. But they loved once, and these belong to
her. Do you suppose they would be a comfort or a curse to her? If—if
—” the connoisseur evidently found difficulty in stating his position.
“Of course I do not mean to say that I believe one of the stones
waned while the other grew more brilliant. I simply say nothing of it;
but I know that he believed it, and I, even I, feel a superstition about
it. I do not want the light in that stone to go out; or if it should, or
could, I do not want to see it. And, besides, if I were a woman, and
that man had loved me so, I should wish those opals.” Here Hayden
looked up and caught Langworthy’s amused, tolerant smile. He
stopped, and there was almost a flush upon his cheek.
“You think I am maudlin—doting—I see,” he said. “Langworthy, I
do hope the Lord will kindly let you die in the harness. You haven’t
any taste for these innocent, green pastures where we old fellows
must disport ourselves, if we disport at all. Now, I want to know if it
would be—er—indelicate to attempt to find out who she is, and to
restore the stones to her?”
Langworthy, who had preserved throughout his usual air of strict
scientific attention, jumped up and began to pace the room.
“His name?” he said.
Hayden gave it.
“I know the man,” said Langworthy, almost reluctantly. “Did any
one who ever saw him forget him? He was on the verge of
melancholia, but what a mind he had!”
“How did you know him, Langworthy?” asked Hayden, with
pathetic eagerness.
“As a patient. It’s a sad story. You won’t like it. You had better keep
your fancies without the addition of any of the facts.”
“Go on,” said Hayden, briefly.
“They live here, you know. He was the only son. He unconsciously
acquired the morphine habit from taking quantities of the stuff for
neuralgic symptoms during a severe protracted illness. After he got
better, and found what had happened to him, he came to me. I had
to tell him he would die if he didn’t break it off, and would probably
die if he did. ‘Oh, no matter,’ he said. ‘What disgusts me is the idea
that it has taken such hold of me.’ He did break it off directly and
absolutely. I never knew but one other man who did that thing. But
between the pain and the shock from the sudden cessation of the
drug, his mind was unbalanced for awhile. Of course the girl’s
parents broke off the engagement. I knew they were travelling with
him last summer. It was a trying case, and the way he accepted his
own weakness touched me. At his own request he carried no money
with him. It was a temptation when he wanted the drug, you see. It
must have been at some such moment, when he contemplated
giving up the struggle, that you met him in the pawn-shop.”
“I am glad I knew enough to respect him even there,” murmured
Hayden, in his beard.
“Oh, you may respect him, and love him if you like. He died a
moral hero, if a mental and physical wreck. That is as good a way as
any, or ought to be, to enter another life—if there is another life.”
“And the woman?” asked the connoisseur.
“Keep the opals, Hayden; they and he are more to you than to her.
She—in fact it is very soon—is to marry another man.”
“Who is—”
“A gilded cad. That’s all.”
Langworthy took out his watch and looked at it. I turned to the
table. What had happened to the dreaming stones? Did a light flash
across from one to the other, or did my eyes deceive me? I looked
down, not trusting what I saw. One opal lay as pale, as pure, as
lifeless, as a moon-stone is. The other glowed with a yet fierier
spark; instead of coming from within, the color seemed to play over
its surface in unrestricted flame.
“See here!” I said.
Langworthy looked, then turned his head away sharply. The
distaste of the scientific man for the inexplicable and irrational was
very strong within him.
But the old man bent forward, the lamp-light shining on his white
hair, and with a womanish gesture caught the gleaming opal to his
lips.
“A human soul!” he said. “A human soul!”
AN UNEARNED REWARD
It is the very last corner of the world in which you would expect to
find a sermon. Overhead hang the Colorado skies, curtains of
deepest, dullest cobalt, against which the unthreatening white clouds
stand out with a certain solidity, a tangible look seen nowhere else
save in that clear air. All around are the great upland swells of the
mountains, rising endlessly, ridge beyond ridge, like the waves of the
sea. In a hollow beside the glittering track is the one sign of human
existence in sight—the sun-scorched, brown railway station. It is an
insignificant structure planted on a high platform. There is a red tool-
chest standing against the wall; a tin advertisement of somebody’s
yeast-cakes is nailed to the clap-boards; three buffalo hides, with
horns still on them, hang over a beam by the coal-shed, and across
the side of the platform, visible only to those approaching from the
west, is written, in great, black letters:
THE WAGES OF SIN IS DEATH.
This legend had no place there on the September afternoon, some
years ago, when Carroll Forbes stepped off the west-bound express
as it halted a minute at the desolate spot. Because it looked to him
like the loneliest place in all the world the notion seized him
suddenly, as the train drew up beside the high platform, to catch up
his valise and leave the car. He was looking for a lonely place, and
looking helplessly. He snatched at the idea that here might be what
he sought, as a drowning man at the proverbial straw.
When the train had gone on and left him there, already repenting
tremulously of what might prove his disastrous folly, a man, who was
possibly the station agent—if this were indeed a station—came
limping toward him with an inquiring look.
Forbes was a handsome man himself, and thoroughly aware of
the value of beauty as an endowment. He was conscious of a half-
envious pang as he faced the blonde giant halting across the
platform. This was, or had been, a singularly perfect specimen of the
physical man. Over six feet in height, muscular, finely proportioned,
fair-haired and fair-skinned, with a curling, blonde beard, and big,
expressionless blue eyes, he looked as one might who had been
made when the world was young, and there was more room for
mighty men than now.
The slight, olive-skinned young man who faced him was conscious
of the sudden feeling of physical disadvantage that comes upon one
in the presence of imposing natural objects, for the man was as
august in his way as the cliffs and canyons.
“I am a—an artist,” said Carroll Forbes. “Is there any place
hereabouts where I can get my meals and sometimes a bed, while I
am sketching in the mountains?”
The man stared at him.
“Would it have been better if I had said I was a surveyor?” asked
Forbes of his confused inner consciousness.
“We feed folks here sometimes—that is, my wife does. Mebbe you
could have a shake-down in the loft. Or there’s Connor’s ranch off
north a ways. But they don’t care about taking in folks up there.”
“Then, if you would ask your wife?” ventured Forbes, politely. “I
shall not trouble you long,” he added.
“Ellen!”
A woman appeared at the door, then moving slowly forward, stood
at her husband’s side, and the admiration Forbes had felt at the sight
of the man flamed into sudden enthusiasm as he watched the wife.
She was tall, with heavy, black hair, great eyes like unpolished jet,
one of the thick white, smooth, perfectly colorless skins, which
neither the sun nor the wind affect, and clear-cut, perfect features.
Standing so, side by side, the two were singularly well worth looking
at.
“What a regal pair!” was Forbes’s internal comment; and while
they conferred together he watched them idly, wondering what their
history was, for of course they had one. It is safe to affirm that every
human creature cast in the mould of the beautiful has, or is to have,
one.
“She says you c’n stay,” announced the man. “Just put those traps
of yours inside, will you?” and, turning, he limped off the length of the
platform at a call from somebody who had ridden up with jingling
spurs.
Forbes, left to his own devices, picked up his valise, then set it
down again and looked around him helplessly, wondering if there
was a night train by which he could get away from this heaven-
forsaken spot.
“If you want to see where you can sleep,” said a voice at his side,
“I will show you.” It was the woman. She bent as she spoke to pick
up some of his impedimenta, but he hastily forestalled her with a
murmur of deprecation.
She turned and looked at him, and as he met her eyes it occurred
to him that the indifference of her face was the indifference of the
desert—arid and hopeless. The look she gave him was searching
and impersonal; he saw no reason for it, nor for the slow, dark color
that spread over her face, and there was less than no excuse for the
way she set her lips and stretched a peremptory hand, saying, “Give
me those,” in tones that could not be disobeyed. To his own
astonishment he surrendered them, and followed her meekly up a
ladder-like flight of steps to the rough loft over the station. It was
unfinished, but partitioned into two rooms. She opened the door of
one of these apartments, silently set his luggage inside, and
vanished down the stairs.
Forbes sat down on the edge of a broken chair and looked about
him.
“Now, in heaven’s name,” he demanded of the barren walls, “what
have I let myself in for, and why did I do it?”
To this question there seemed no sufficient answer, and for awhile
he sat there fretting with the futile anxiety of a man who knows that
his fate pursues him, who hopes that this turning or that may help
him to evade it, yet always feels the benumbing certainty that the
path he has taken is the shortest road to that he would avoid. When
at last—recognizing that his meditations were unprofitable—he rose
and went down the stairs, it was supper-time.
The woman was uncommunicative, but he could feel that her eyes
were on him. The man—it occurred to Forbes that he had probably
been drinking—was talkative. After the meal was over they went
outside. Forbes, by way of supporting his pretence of being an artist,
took out a pocket sketch-book and made notes of the values of the
clouds and the outlines of the hills against the sky in a sort of artistic
short-hand. The man Wilson sat down on a bench and began to talk.
Between the exciting effects of the whiskey he had taken, the
soothing influence of the cigar Forbes proffered him, and a natural
talent for communicativeness, he presently went on to tell his own
story. Forbes listened attentively. It seemed a part of the melodrama
of the whole situation and was as unreal to him as the flaming
miracle of the western skies or his own presence here.
“So the upshot of it all was that we just skipped out. She ran away
with me.”
It was a curious story. As Forbes listened he became aware that it
was one with which he had occasionally met in the newspapers, but
never in real life before. It was, apparently, the story of a girl
belonging to a family of wealth and possibly of high social traditions
—naturally he did not know what importance to attach to Wilson’s
boast that his wife belonged “to the top of the heap”—who had
eloped with the man who drove her father’s carriage.
The reasons for this revolt against the natural order of her life was
obscure; there was, perhaps, too high a temper on her side and too
strict a restraint on the part of her guardians. There was necessarily
a total absence of knowledge of life; there was also the fact that the
coachman was undoubtedly a fine creature to look at; there might
have been a momentary yielding on the part of a naturally dramatic
temperament to the impulse for the spectacular in her life.
But whatever the reasons, the result was the same. She had
married this man and gone away with him, and they had drifted
westward. And when they had gone so far west that coachmen of his
stamp were no longer in demand, he took to railroading, and from
brakeman became engineer; and finally, being maimed in an
accident in which he had stood by his engine while the fireman
jumped—breaking his neck thereby—he had picked up enough
knowledge of telegraphy to qualify him for this post among the
mountains. He and his handsome wife lived here and shared the
everlasting solitude of the spot together, and occasionally fed stray
travellers like this one who had dropped down on them to-day.
“He drinks over-freely and he swears profusely,” mused Forbes,
scrutinizing him, “but he is too big to be cruel, and he still worships
her beauty as she, perhaps, once worshipped his; and he still feels
an uncouth pride in all that she gave up for his sake.”
It had never occurred to him before to wonder what the after-life of
a girl who eloped with her father’s servant might be like. He
speculated upon it now. By just what process does a woman so
utterly déclassée adjust herself to her altered position? Would she
make it a point to forget, or would every reminder of lives, such as
her own had been, be a turning of the knife in her wound? Would not
a saving recollection of the little refinements of life cling longer to a
weak nature than to a strong one under such circumstances?
This woman apparently gave tongue to no vain regrets, for her
husband was exulting in the “grit” with which she had taken the
fortunes of their life. “No whine about her,” was his way of expressing
his conviction that the courage of the thoroughbred was in her.
“No, sir; there’s no whine about her. Un she’s never been sorry,
un, s’help me, she sha’n’t never be,” concluded Wilson. There were
maudlin tears in his eyes.
“Few men can say that of their wives,” said Forbes’s smooth,
sympathetic voice. “You are indeed fortunate.”
While her husband was repeating the oft-told tale of their conjugal
happiness, Ellen Wilson had done her after-supper work, and,
slipping out of the door, climbed the short, rocky spur to the north of
the station. Beyond the summit, completely out of sight and hearing,
there was a little hollow that knew her well, but never had it seen her
as it saw her now, when, throwing herself down, her face to the
earth, she shed the most scalding tears of all her wretched years.
They were such little things this stranger had done—things so
slight, so involuntary, so unconscious that they did not deserve the
name of courtesies, but they were enough to open the flood-gates of
an embittered heart. There was a world where all the men were
deferential and all the women’s lives were wrapped about with the
fine, small courtesies of life—formal, but not meaningless. It had
been her world once and now was so no longer.
Good or bad, she knew little and cared less, this man had come
from that lost world of hers, as she was made aware by a thousand
small signs, whose very existence she had forgotten; and silently,
fiercely she claimed him as an equal.
“I—I too was—” Slow tears drowned the rest.
She could have told him how a déclassée grows used to it. She
knew how the mind can adjust itself to any phase of experience, and
had learned that what woman has undergone, woman can undergo
—yes, and be strong about it. She knew how, under the impulse of
necessity, the once impossible grows to be the accepted life, and the
food that could not be swallowed becomes the daily bread.
When the struggle for existence becomes a hand-to-hand fight,
traditions of one’s ancestry do not matter, except, possibly, that some
traditions bind you to strength and silence, while others leave you
free to scream. She knew what it was to forget the past and ignore
the future, and survey the present with the single-hearted purpose of
securing three meals a day, if possible; two, if it were not.
She had forgotten with what facility she might the faces and
scenes that once were dear to her. She had nothing to do with them
any longer, as she knew. She might, perhaps, have heard their
names without emotion. But, even in this day and generation and
among this democratic people, in the soul of a woman bred as she
had been the feeling for her caste is the last feeling that dies. And to
her anguish she found that in her it was not yet dead.
The color died from the sky, and the stars came swiftly out.

Development in Wastewater Treatment Research and Processes: Microbial Degradation of Xenobiotics Through Bacterial and Fungal Approach Maulin P. Shah

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Development in Wastewater Treatment Research and Processes: Microbial Degradation of Xenobiotics Through Bacterial and Fungal Approach Maulin P. Shah

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Development in Wastewater Treatment

Research and Processes: Microbial

Copyright © 2022 Elsevier Inc. All rights reserved.

Library of Congress Cataloging-in-Publication Data

For information on all Elsevier publications visit our

Publisher: Susan Dennis

Typeset by TNQ Technologies

CHAPTER 1 Microbial degradation of xenobiotics in bioelectrochemical

2.7.2 Temperature ..................................................................................................33

5.3.2 Recommendations ........................................................................................ 111

7.3.7 Lead ............................................................................................................ 142

9.8 Bioleaching (biomining).......................................................................................186

CHAPTER 11 Microbial-mediated explosives removal and its impact

CHAPTER 13 Counterbalancing common explosive pollutants (TNT, RDX,

14.5.6 Enzymes-based biodegradation of various industrial pollutants ...........324

16.6.1 Nanozymes ...............................................................................................371

18.2.3 Asphaltenes and waxes.............................................................................413

20.3 Oil bioremediation................................................................................................455

Index ................................................................................................................................................. 467

Luciana Porto de Souza Vandenberghe

1.1 Introduction to xenobiotics

Development in Wastewater Treatment Research and Processes. https://doi.org/10.1016/B978-0-323-85839-7.00020-7 1

1.2 Conventional processes of xenobiotics degradation and

1.2.2 Membrane processes

1.2.3 Advanced oxidation processes

removal of antibiotics from wastewater. Ozone at high pH increases the production

1.2.4 Biological treatment processes

processes including conventional activated sludge, biological nutrient removal, and

1.3 Xenobiotics degradation in bioelectrochemical

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